Fluorescein transport in isolated proximal tubules in vitro: epifluorometric analysis.
An epifluorometric method was used to quantify the bidirectional fluxes of fluorescein across the basolateral surfaces of nonperfused rabbit tubule segments in vitro. Proximal S2 segments, but not cortical collecting tubules or cortical thick ascending limbs, accumulated fluorescein to levels in cytoplasm over 100-fold greater than in the external medium. The rate of intracellular fluorescein accumulation was dependent on the concentration of the ligand in the external bath. The apparent Km was 10 microM and the Vmax was 623 x 10(-6) mol.min-1.l-1. Probenecid and ouabain inhibited fluorescein accumulation. We conclude that fluorescein is transported into the cytoplasm of proximal tubules by basolateral mechanisms that share features in common with the classical organic anion system. This fluorescent compound offers some unique advantages for the study of the organic anion transport system in intact cells.
Sullivan
L P
LP
Department of Physiology, University of Kansas School of Medicine, Kansas City 66103.
Grantham
J A
JA
Rome
L
L
Wallace
D
D
Grantham
J J
JJ
D000818:Animals; D001692:Biological Transport; D005260:Female; D019793:Fluorescein; D005452:Fluoresceins / Q000378:metabolism*; D005453:Fluorescence; D005470:Fluorometry / Q000379:methods*; D066298:In Vitro Techniques; D007687:Kidney Tubules, Proximal / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D008027:Light; D009994:Osmolar Concentration; D011339:Probenecid / Q000494:pharmacology; D011817:Rabbits; D013997:Time Factors
D005452:Fluoresceins; D011339:Probenecid; D019793:Fluorescein